ŞAHİN DİREKEL, ÜLKÜ KARAMAN, SEDA TEZCAN ÜLGER, SEMRA UTKU, GÖNÜL ASLAN, MEHTAP UYSAL, M. Fethi ŞAHİN, Nuran DELİALİOĞLU, MAHMUT ÜLGER, Gürol EMEKDAŞ

Hidrazon Yapısındaki On Farklı Bileşiğin Antileishmanial Aktivitesinin Araştırılması

Leishmaniasis Türkiye ve dünyada önemli paraziter bir hastalıktır. Leishmaniasis tedavisinde sodyum stibogluconate, miltefosin, paramomisin, amfoterisin B ve pentamidin gibi anti-leishmanial ilaçlar kullanılmaktadır. Fakat leishmaniasisin kemoterapisinde kullanılan bu ilaçların nefrotoksik, hepatotoksik ve teratojenik yan etkileri görülebilmektedir. Ayrıca antimon bileşiklerine karşı direnç gelişmesi nedeniyle de yeni terapötik ajanların keşfedilmesi ve geliştirilmesine öncelik verilmesi gerektiği düşünülmüştür. Bu çalışmanın amacı, sentezlenmiş hidrazon yapısındaki on farklı bileşiğin (5a-5j) Leishmania infantum promastigotlarına karşı anti-leishmanial aktivitesinin mikrodilüsyon yöntemiyle belirlenmesidir. Hazırlanan hidrazon bileşikleri, konsantrasyonu 6 µg/ml olacak şekilde RPMI-1640 besiyerlerine eklenmiş ve bileşiklerin mikroplakta konsantrasyon aralığı 3 - 0.003 µg/ml olacak şekilde seri dilüsyonları yapılmıştır. Mikroplaklarda mikrodilüsyon besiyeri yöntemi uygulanarak, üzerine hemositometrede hücre sayısı 2.5x107hücre/ml olacak şekilde ayarlanmış standart Leishmania infantum promastigotları eklenerek 27°C'de inkübe edilmiştir. Yirmi saat sonra mikroplakların üzerine alamar mavisi eklenerek 4 saat inkübe edilmiştir. Promastigotların üremesi 24, 48 ve 72. saatlerde değerlendirilmiştir. Kuyucuklarda rengin maviden pembeye dönmesi parazitin ürediği, rengin değişmeden kalması ise parazitin üremediği şeklinde değerlendirilmiştir. Bu çalışmada, Leishmania infantum promastigotlarına karşı en etkili maddelerin 5e, 5g ve 5h bileşikleri (MİK 0.187 µg/ml) olduğu, en etkisiz bileşiğin ise 5i bileşiği (MİK 3 µg/ml) olduğu saptanmıştır. Sentezlenen bileşiklerin ilaç olarak kullanılabilmesi için; gerekli olduğu düşünülen in vitro makrofaj kültüründe Leishmania amastigotlarına karşı etkinliği ve in vivo olarak deneysel hayvan modellerinde kontrol çalışmalarına ihtiyaç vardır.

Investigation of Anti-leishmanial Activity of the Ten Different Hydrazone Derivatives

Leishmaniasis is an important parasitic disease in Turkey and the world. Anti-leishmanial drugs such as sodium stibogluconate, miltefosine, paramomycin, amphotericin B, and pentamidine are used for the treatment of leishmaniasis. However, the drugs, used for the chemotheraphy of leishmaniasis, have some side effects such as nephrotoxicity, hepatotoxicity, and teratogenicity. In addition, it is deemed that the discovery and development of the new therapeutic agents must be given priority due to the development of resistance against antimony compounds. The purpose of this study is detecting the anti-leishmanial activity of ten different synthesized hydrazone compounds against Leishmania infantum promastigotes via the microdilution method. The prepared hydrazone compounds, having the concentration of 6 µg/ml, were added to RPMI-1640 media and the dilution of the hydrazone derivates was performed in the wells of microplates in the range of concentrations of 3 - 0.003 µg/ml. The microdilution broth method in the microplates was prepared, than the adjusted standard Leishmania infantum promastigotes, 2.5x107cells/mL, were added, into the mentioned microplates which was incubated in 27°C. Twenty h later, the alamar blue were added on the microplates and they were incubated for 4 h. The proliferation of promastigotes was evaluated in 24, 48, and 72 h. It was considered that changing the color from blue to pink in the wells were exhibiting the growth of parasites, while the unchanged color was not. The present study has revealed that the most effective substances against Leishmania infantum promastigotes were 5e, 5g, and 5h compounds (MIC 0.187 µg/ml) while the least effective compound was 5i (MIC 3 µg/ml). There is a need for further studies on in vitro activity against the Leishmania amastigotes in macrophages cultures and in vivo experimental animal models for the synthesized compounds showing anti-leishmanial effect in the present study.

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